Part:BBa_K1463560

Reverse RBS

Reverse ribosome binding site designed using the Salis ribosome binding site calculator to have a strength of 12,204 when placed downstream of the reversed rfp part BBa_K1463520

Usage and Biology

This has been used to drive expression of the reversed RFP part BBa_K1463520 in our phiC31 integrase-based inversion switch BBa_K1463000.

It gave a higher level of fluorescence than a previous design for a reverse RBS BBa_I742130

Fig. 1: Red fluorescence of colonies containing BBa_I742130 or K1463560 reversed RBSs downstream of rfp gene transcribed from the PhiC31 integrase switch K1463000

It was designed using the Salis RBS calculator software using the RBS sequence NNNNNNNNNNCTCTAGTA (10 Ns followed by a reversed scar) upstream of the RFP open reading frame.

Fig. 2: Salis RBS calculator output for reverse RBS design

As can be seen, the designed RBS gives the start codon at position 39 a calculated translation initiation rate of "12204.43". This is 25 fold higher than the calculated translation initiation rate of BBa_I742130.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]